Process for producing monoclonal antibodies reactive with human breast cancer

Abstract

Monoclonal antibodies demonstrating a reactivity with human breast cancer are produced. The hybridoma cultures secreting immunoglobins are produced by hydridoma technology. Splenic lymphocytes of mice, immunized with membrane-enriched fractions of metastatic human mammary carcinoma tissue are fused with the NS-1 non-immunoglobulin-secreting murine myeloma cell line. Screening of immunoglobulin reactivities and double cloning of cultures yielded 11 monoclonal antibodies that demonstrated activities with the surface of human mammary tumor cells and not with the surface of apparently normal human tissues. These monoclonal antibodies aid in the diagnosis, prognosis and treatment of human breast cancer.

Patent number: 4522918
Filing date: Dec 15, 1981
Issue date: Jun 11, 1985
Inventors: Jeffery Schlom, David Colcher, Marianna Nuti, Patricia H. Hand, Faye Austin
Primary Examiner: John Edward Tarcza

download


What is claimed is:

1. A process for producing antibodies from hyridoma cultures comprising the steps of

(1) taking splenic lymphocytes of mice previously immunized with membrane-enriched fractions of immunoglobulin-depleted cancer cells;

(2) fusing the lymphocytes with murine myeloma cell line, and

(3) culturing the cell line in an in vitro culture medium or in vivo therefor to produce antibodies, which antibody:

(a) reacts and binds with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts;

(b) reacts and binds with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and

(c) does not react with normal cells derived from breast, skin, lung, bone marrow, kidney, spleen and uterus.

2. The method according to claim 1 wherein said human cancer cells are from metastatic mammary carcinoma in the liver.

3. The method according to claim 1 wherein the depletion is performed using affinity chromatography.

4. The method according to claim 1 wherein the depletion is performed using protein A bound to a solid matrix.

5. The method according to claim 4 wherein said matrix is Sepharose.

6. A process of claim 1 wherein the murine myeloma cell line is non-immunoglobulin secreting murine myeloma cell line designated as NS-1.

7. A method for preparing monoclonal antibody which antibody:

(a) reacts and binds with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts;

(b) reacts and binds with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and

(c) does not react with normal cells derived from breast, skin, lung, bone marrow, kidney, spleen and uterus, which comprises the steps of:

(1) immunizing mice with membrane enriched fractions of immunoglobulin-depleted cancer cells;

(2) removing the spleens from said mice and making a suspension of spleen cells;

(3) fusing said spleen cells with NS-1 non-immunoglobulin-secreting murine myeloma cell line in the presence of a fusion promoter;

(4) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused cells;

(5) evaluating the supernatant in each well containing a hybridoma for the presence of the desired antibody;

(6) selecting and cloning hybridoma producing the desired antibody; and

(7) recovering the antibody from supernatant above said clones.

Early detection of breast cancer using transillumination

Abstract

Several methods have been developed for improving transillumination devices such that they may be effectively used for breast cancer examinations. Each of the methods involves a particluar technique for reducing scattered light. If light which passes straight through a breast can be distinguished from light which is scattered within the breast, better images can be produced. In addition, biochemical markers have been developed which associate with cancerous tissue and enhance the contrast by absorbing light of specific wavelengths.

Patent number: 4945239
Filing date: Mar 29, 1989
Issue date: Jul 31, 1990
Inventors: Abund O. Wist, Ramendra N. Pandey, Panos P. Fatouros
Assignee: Center for Innovative Technology

download


What is claimed is:

1. A transillumination apparatus for the early detection of breast caner, comprising:

a source of nonionizing radiation for producing radiation in a specific frequency range, said nonionizing radiation having an intensity which permits passing through a breast;

an optical receiver for receiving radiation from said source of nonionizing radiation which passes through said breast; and

a biochemical marker associated with cancer cells within said breast which absorbs radiation in said specific frequency range.

2. A transillumination apparatus for the early detection of breast cancer as recited in claim 1 where in said biochemical marker is fluorescein isothiocyanate covalently bonded with human transferrin.

3. A transillumination apparatus for the early detection of breast cancer as recited in claim 1 further comprising collimating means for collimating said radiation produced by said source of non-ionizing radiation into a collimated beam of radiation and scanning means for scanning said collimated beam over said breast.

4. A transillumination apparatus as recited in claim 3 further comprising a pair of pin hole boxes positioned on opposite sides of said breast, a first of said pair of pin hole boxes being positioned between said source of radiation and said breast, a second of said pair of pin hole boxes being positioned between said breast and said optical receiver, each of said pin hole boxes having apertures which permit said collimated beam to pass therethrough.

5. A transillumination apparatus as recited in claim 3 further comprising a pair of matched polarizing filters positioned on opposites sides of said breast, a first of said pair being positioned between said source of radiation and said breast, a second of said pair being positioned between said breast and said optical receiver, said first of said pair of polarizing filters permitting only radiation having a specific plane of polarization to pass through said breast, said second of said pair of polarizing filters permitting only radiation having said specific plane of polarization to impinge on said optical receiver.

6. A transillumination apparatus as recited in claim 3 further comprising a phase conjugated mirror for returning radiation of the same phase as said radiation produced by said source, said collimated beam impinging on said phase conjugated mirror before said returning radiation impinges on said optical receiver.

7. A transillumination apparatus as recited in claim 3 wherein said optical receiver comprises a one or more photodetectors, each of said photodetectors producing an electrical signal which corresponds to the intensity of radiation received after having passed straight through said breast.

8. A transillumination apparatus as recited in claim 7 further comprising compressing means for compressing said electrical signal to a compressed signal having a frequency range compatible with photographic film and exposing means for exposing said photographic film to light having an intensity corresponding to said compressed signal.

9. A transillumination apparatus as recited in claim 8 wherein said exposing means comprises an adapter for receiving said compressed signals and a light emitting diode connected to said adapter, said light emitting diode emitting said light having said intensity corresponding to said compressed signal.

10. A transillumination apparatus as recited in claim 3 wherein said optical receiver comprises photographic film, said photographic film being exposed by non-ionizing radiation which has passed straight through said breast.

11. A transillumination apparatus as recited in claim 3 wherein said optical reservoir comprises a photodetector array having a plurality of photodetectors arranged to receive said collimated beam, and further comprising determining means for determining which photodetector of said plurality of photodetectors said collimated beam is directed towards by said scanning means, activating means for selectively activating said photodetector to receive radiation passing straight through said breast, and displaying means for displaying an image of said breast, said photodetector producing an electrical signal proportional to the intensity of said radiation which passes straight through said breast, said displaying means using said electrical signal to present said image of said breast.

12. A transillumination apparatus as recited in claim 3 further comprising a pair of light shutters and controlling means for controlling the opening and closing of said pair of light shutters, a first light shutter being positioned between said source of non-ionizing radiation and said breast, a second light shutter being positioned between said breast and said optical receiver, said controlling means opening said first light shutter for a first short time period and then closing said first light shutter, said controlling means opening said second light shutter for a second short time period and then closing said second light shutter, said first short time period occurring at a predetermined time interval before said second short time period.

13. A transillumination apparatus as recited in claim 12 wherein said predetermined time interval is equivalent to the time required for said non-ionizing radiation to pass from said first light shutter through said breast to said second light shutter.

14. A transillumination apparatus as recited in claim 13 wherein said controlling means provides pulsed signals to said first and second light shutters for opening and closing said first and second light shutters.

15. A transillumination apparatus as recited in claim 13 wherein said predetermined timed interval is determined from standards which simulate breast tissue.

16. A transillumination apparatus for the early detection of breast cancer as recited in claim 1 wherein said source emits a divergent beam of radiation and said optical receiver comprises a photodetector array having a plurality of photodetectors, and further comprising a pulsing means for pulsing said source of non-ionizing radiation to emit pulses of radiation and distinguishing means for distinguishing first arriving radiation from later arriving radiation at each of said photodetectors in said array, said first arriving radiation being the radiation which arrives at said photodetector first after said pulse and said later arriving radiation being the radiation which arrives at said photodetector after said first arriving radiation.

17. A transillumination apparatus for the early detection of breast cancer as recited in claim 1 wherein said source is a laser diode which emits a plurality of collimated beams of radiation and said optical receiver comprises a photodetector array having a plurality of photodetectors, each of said collimated beams from said laser diode being aimed at specific photodetectors in said array.

18. A transillumination apparatus as recited in claim 1 wherein said source emits a divergent beam of radiation and said optical receiver comprises a photodetector array having a plurality of photodetectors, and further comprising pulsing means for pulsing said source to emit pulses of non-ionizing radiation and activating means for selectively activating each of said photodetectors in said array to receive radiation which has passed straight through said breast.

19. A transillumination apparatus as recited in claim 1 wherein said source is a laser diode which emits a plurality of collimated beams of radiation and said optical receiver comprises a photodetector array having a plurality of photodetectors, and further comprising pulsing means for pulsing said laser diode to emit pulses of non-ionizing radiation and activating means for selectively activating each of said photodetectors in said array to receive radiation which has passed straight through said breast.

20. A transillumination apparatus as recited in claim 1 wherein said optical receiver comprises a photodetector array with a plurality of photodetectors, and further comprising first and second phase plates, said first phase plate being positioned between said source and said breast, said second phase plate being positioned between said breast and said photoedetector array, said first phase plate polarizing said non-ionizing radiation before it impinges on said breast, said second phase plate permitting only light having a plane of polarization matched to the light impinging on said breast to pass therethrough to said photodetector array.

21. A transillumination apparatus as recited in claim 20 wherein said first and second phase plates are identical, each of said phase plates having a plurality of sections, each of said sections being ninety degrees out of phase with adjacent sections.

22. A transillumination apparatus as recited in claim 20 wherein said first and second phase plates are not identical, each of said phase plates having a plurality of sections, each of said sections being ninety degrees out of phase with adjacent sections.

23. A transillumination apparatus as recited in claim 1 wherein said optical receiver comprises photographic film, and further comprising a semi-permeable mirror positioned between said source of non-ionizing radiation and said breast, said semi-permeable mirror dividing said non-ionizing radiation into a sample beam and a reference beam, said sample beam being directed through said breast before impinging on said photographic film, said reference beam being directed to said photographic film without passing through said breast.

24. A transillumination apparatus for the early detection of breast caner, comprising:

a source of non-ionizing radiation for producing nonionizing radiation of sufficient intensity to pass through a breast;

an optical receiver for receiving radiation from said source of nonionizing radiation which passes through said breast;

a first polarizing means positioned between said source and said breast for polarizing said nonionizing radiation before it impinges on said breast; and

a second polarizing means matched with said first polarizing means positioned between said breast and said optical receiver for permitting only radiation having a polarity matched to the radiation produced by said first polarizing means to impinge on said optical receiver.

25. A transillumination apparatus as recited in claim 24 wherein said source of non-ionizing radiation produces a collimated beam of radiation which is scanned over said breast, said optical receiver detecting said collimated beam as it is scanned over said breast.

26. A transillumination apparatus as recited in claim 25 wherein said optical receiver comprises a photodetector array with a plurality of photodetectors, each of said photodetectors producing an electrical signal which corresponds to the intensity of received radiation.

27. A transillumination apparatus as recited in claim 25 wherein said optical receiver comprises photographic film, said photographic film being exposed by non-ionizing radiation which has passed straight through the breast.

28. A transillumination apparatus for the early detection of breast caner, comprising:

a source of non-ionizing radiation for producing nonionizing radiation of sufficient intensity to pass through a breast;

an optical receiver for receiving radiation from said source of nonionizing radiation which passes through said breast; and

first and second phase plates having a plurality of polarized sections with adjacent sections being out of phase with respect to one another, said first phase plate being positioned between said source and said breast, said second phase plate being positioned between said breast and said optical receiver.

29. A transillumination apparatus as recited in claim 28 wherein said first and second phase plates are identical.

30. A transillumination apparatus as recited in claim 28 wherein said first and second phase plates are not identical.

31. A transillumination apparatus for the early detection of breast cancer, comprising:

a source of non-ionizing radiation for producing a collimated beam of radiation of sufficient intensity to pass through a breast;

scanning means for scanning said collimated beam of radiation over said breast;

a phase conjugated mirror for receiving radiation which has passed through the breast, said phase conjugated mirror returning radiation in the same phase as said radiation in said collimated beam; and

an optical receiver for receiving returning radiation from said phase conjugated mirror.

32. A transillumination apparatus for the early detection of breast cancer, comprising:

a source of non-ionizing radiation for producing nonionizing radiation of sufficient intensity to pass through a breast;

an optical receiver for receiving radiation which passes through said breast;

pulsing means for providing pulses of said nonionizing radiation; and

discriminating means for discriminating between radiation which passes straight through said breast and radiation which is scattered within said breast based on the time required for a pulse of said nonionizing radiation to traverse said breast.

33. A transillumination apparatus as recited in claim 32 wherein said optical receiver comprises a photodetector array with a plurality of photodetectors, each of said plurality of photodetectors being selectively activated by said discriminating means to receive radiation at a specific time interval after a pulse of said nonionizing radiation is emitted from said source, said specific time interval corresponding to the time required for said radiation to pass through said breast.

34. A transillumination apparatus as recited in claim 33 wherein said specific time interval is determined relative to standards simulating breast tissue.

35. A transillumination apparatus as recited in claim 32 further comprising first and second phase plates positioned on opposite sides of said breast, each of said phase plates having a plurality of polarized sections where adjacent sections are out of phase with respect to each other.

36. A transillumination apparatus for the early detection of breast cancer, comprising:

a light source producing a collimated beam of non-ionizing radiation of sufficient intensity to pass through a breast;

an optical receiver for receiving radiation which has passed straight through said breast;

first and second light shutters positioned on opposite sides of said breast in the path of said collimated beam; and

a controller for opening and closing said first and second light shutters on a timed basis.

37. A transillumination apparatus as recited in claim 36 wherein said first and second light shutters are electronically actuated by pulses produced by said controller.

38. A transillumination apparatus as recited in claim 36 wherein said optical receiver comprises photographic film which is exposed by light allowed through said second shutter.

39. A transillumination apparatus for the early detection of breast cancer, comprising:

a source of non-ionizing radiation for producing nonionizing radiation of sufficient intensity to pass through a breast;

a photodetector array comprised of a plurality of photodetectors, each of said photodetectors receiving radiation which has passed through said breast, each of said photodetectors producing an electrical signal which corresponds to the intensity of radiation received;

compressing means for compressing said electrical signal to a compressed signal having a frequency range compatible with photographic film; and

exposing means for exposing said photographic film to light having an intensity corresponding to said compressed signal.

40. A transillumination apparatus as recited in claim 39 wherein said exposing means comprises an adapter for receiving said compressed signals and a light emitting diode connected to said adapter, said light emitting diode emitting said light having said intensity corresponding to said compressed signal.

41. A transillumination apparatus for the early detection of breast cancer, comprising:

a source of non-ionizing radiation for producing nonionizing radiation of sufficient intensity to pass through a breast;

photographic film for receiving radiation from said source of nonionizing radiation; and

a semi-permeable mirror positioned between said source of non-ionizing radiation and said breast, said semi-permeable mirror dividing said non-ionizing radiation into a sample beam and a reference beam, said sample beam being directed through said breast before impinging on said photographic film, said reference beam being directed to said photographic film without passing through said breast, whereby a hologram image is created from the interference of said sample beam with said reference beam.

42. A method for providing a collimated beam of light with a very narrow diameter, comprising the steps of:

directing a beam of light through a first plate with a first aperture, said first aperture having a diameter on the order of wavelengths, said first aperture allowing a collimated beam of light having a diameter of said first aperture to pass straight through and refracting the remaining light from said beam of light; and

providing a second plate with a second aperture in the path of said collimated beam of light, said second aperture being slightly larger in diameter than said first aperture, said second plate allowing said collimated beam of light having said diameter equivalent to said first aperture to pass through said second aperture and preventing refracted light from passing therethrough.

43. A method for detecting breast cancer using a transillumination apparatus, comprising the steps of:

scanning a collimated beam of electromagnetic radiation of a first wavelength which is strongly absorbed by fat over a breast;

detecting radiation of said first wavelength which has passed straight through said breast;

preventing scattered light of said first wavelength from being detected;

creating a fat line image from the detected radiation of said first wavelength;

scanning a collimated beam of electromagnetic radiation of a second wavelength which is strongly absorbed by water over said breast;

detecting radiation of said second wavelength which has passed straight through the breast;

preventing scattered light of said second wavelength from being detected;

creating a water line image from the detected radiation of said second wavelength; and

comparing said fat line image and said water line image, tumors being detected by an increased absorption in said water line image with a corresponding decrease in said fat line image.

44. A method as recited in claim 43 further comprising the steps of:

scanning a collimated beam of electromagnetic radiation of a third wavelength which is strongly absorbed by breast tissue over said breast;

detecting radiation of said third wavelength which has passed straight through said breast;

preventing scattered radiation of said third wavelength from being detected;

creating an overall absorption image from the detected radiation of said third wavelength; and

superimposing the fat line image, the water line image and the overall absorption image to create composite image of the breast which shows the relative location of tumors within said breast.

45. A method for creating a composite image of a breast for the early detection of cancer, comprising the steps of:

creating an overall absorption image of the breast by imaging the breast at a first wavelength;

creating a lesion specific image of said breast by imaging the breast at a second wavelength which is specific to a biochemical marker which associates itself with a breast lesion; and

superimposing the lesion specific image on the overall absorption image to create a composite image of the breast.

46. A method for locating a tumor within a breast, comprising the steps of:

imaging said breast at a first angle to create a first image, lesions in said breast appearing on said first image at a first location;

imaging said breast at a second angle to create a second image, lesions in said breast appearing on said second image at a second location; and

determining the three dimensional location of a lesion within the breast from the position of said first location on said first image and the position of the second location on said second image.

47. A method of preventing scattered light from being detected by a photodetector array positioned under a breast when said breast is imaged with a source of non-ionizing radiation, comprising the steps of:

pulsing said source of non-ionizing radiation such that pulses of light impinge on said breast;

selectively activating each of said photodetectors in said photodetector array to receive light which passes straight through said breast at a first programmed time period after a pulse of radiation is emitted from said source, said first programmed time period being equivalent to the time required for a pulse of light to traverse said breast; and

de-activating said each of said photodetectors in said photodetector array such that they cannot receive light at a second programmed time period, said second programmed time period corresponding to a time in which scattered light traverses said breast.

48. A method for the early detection of breast using a transillumination apparatus, comprising the steps of:

opening and closing a first shutter in a path of a collimated beam of non-ionizing radiation emitted from a source of non-ionizing radiation, said non-ionizing radiation being of sufficient intensity to pass through a breast under examination, said first shutter being positioned between said source and said breast;

opening and closing a second shutter in said path of said collimated beam, said second shutter being positioned between said breast and a photodetector array which detects radiation which passes through said second shutter; and

controlling the time of openings and closings of said first and second shutters.

49. A transillumination apparatus for the early detection of breast cancer, comprising:

a source of non-ionizing radiation for producing a collimated beam of nonionizing radiation of sufficient intensity to pass through a breast;

an optical receiver for receiving radiation from said source of nonionizing radiation which passes through said breast;

a modifying means for modifying radiation in said collimated beam to include identifying information, said modifying means modifying said collimated beam before it passes through said breast; and

a discriminating means, positioned between said breast and said optical receiver, for allowing only radiation including said identifying information to impinge on said optical receiver.

50. A method of preventing scattered light produced in a breast that is impinged with a collimated beam of light from being detected by a photodetector array, comprising the steps of:

modifying said collimated beam of radiation to include identifying information before it passes through said breast; and

allowing only radiation including said identifying information to be detected by said photodetector array.

Monoclonal antibodies reactive with human breast cancer

Abstract

Monoclonal antibodies demonstrating a reactivity with human breast cancer are produced. The hybridoma cultures secreting immunoglobins are produced by hydridoma technology. Splenic lymphocytes of mice, immunized with membrane-enriched fractions of metastatic human mammary carcinoma tissue are fused with the NS-1 non-immunoglobulin-secreting murine myeloma cell line. Screening of immunoglobulin reactivities and double cloning of cultures yielded 11 monoclonal antibodies that demonstrated activities with the surface of human mammary tumor cells and not with the surface of apparently normal human tissues. These monoclonal antibodies aid in the diagnosis, prognosis and treatment of human breast cancer.

Patent number: 4612282
Filing date: Mar 1, 1985
Issue date: Sep 16, 1986
Inventors: Jeffrey Schlom, David Colcher, Marianna Nuti, Patricia H. Hand, Faye Austin
Assignee: The United States of America as represented by the Secretary of the Department of Health and Human Services
Primary Examiner: John Edward Tarcza

download


What is claimed is:

1. Antibodies from hybridoma cultures produced by the steps of

(1) taking splenic lymphocytes of mice previously immunized with membrane-enriched fractions of immunoglobulin-depleted cancer cells;

(2) fusing the lymphocytes with a myeloma cell line, and

(3) culturing the hybridoma cell line in an in vitro culture medium or in vivo therefor to produce antibodies, which antibodies are selected from one member of the group consisting B6.2, B14.2, B39.1, F64.5,B25.2, B84.1, B38.1, B50.4 and B50.1 and which

(a) react and bind with extracts from human metastatic mammary carcinoma cells from involved livers but not with liver cell extracts;

(b) react and bind with at least one of the mammary carcinoma cell lines, BT-20, MCF-7, ZR-75-1, but not with lung, vulva epidermoid or oral epidermoid and not with rhabdomyosarcoma, fibrosarcoma and melanoma; and

(c) do not react with normal cell derived from breast, skin, lung, bone marrow, kidney, spleen and uterus.

2. Antibodies selected from one member of the group consisting of B6.2, B25.2, B72.3, F25.2, B38.1, and B50.4.

3. Antibodies selected from one member of the group consisting of B6.2, B25.2, B38.1, F25.2, B72.3 and B50.4, which bind human mammary carcinoma lines designated BT-20, MCF-7 and ZR-75-1.

4. A mouse myeloma cell line selected from the group consisting of ATCC #HB8106, #HB8107, #HB8108, #HB8109, #HB8110, and #HB8111.

5. A method of detecting a small number of mammary cancer cells in micro-lesions in thin tissue sections or body fluids by applying monoclonal antibodies selected from the group consisting of monoclonal antibodies designated B6.2, B25.2, B72.3, F25.2, B38.1, and B50.4 to the tissue, adding quantity of goat (anti-mouse Ig) antibody conjugated with peroxidase and fixing with diaminobenzidene and peroxide and staining with hemotoxylin, and after fixing, examining for reddish brown colored cells indicative of mammary cancer cells.

6. In an assay to detect mammary carcinoma cells involving contacting a sample with an antibody under conditions which allow the formation of an antibody antigen complex and measurement of the complex formation, the improvement comprises using monoclonal antibodies selected from the group consisting of B6.2, B25.2, B72.3, F25.2, B38.1, and B50.4

Monoclonal anti-human breast cancer antibodies

bstract
Murine monoclonal antibodies are prepared and characterized which bind selectively to human breast cancer cells, are IgGs or IgMs, and when conjugated to ricin A chain, exhibit a TCID 50% against at least one of MCF-7, CAMA-1, SKBR-3, or BT-20 cells of less than about 10 nM. Methods for diagnosing, monitoring, and treating human breast cancer with the antibodies or immunotoxins made therefrom are described.

Patent number: 4753894
Filing date: Jan 11, 1985
Issue date: Jun 28, 1988
Inventors: Arthur E. Frankel, David B. Ring, Michael J. Bjorn
Assignee: Cetus Corporation

download

What is claimed is:

1. A murine monoclonal antibody that:

(a) binds selectively to human breast cancer cells;

(b) has a G or M isotype;

(c) when conjugated to ricin A chain, exhibits a TCID 50% of less than about 10 nM against at least one of MCF-7, CAMA-1, SKBR-3, or BT-20 cells; and

(d) binds a human breast cancer antigen that is also bound by a reference antibody selected from the group consisting of 260F9, 113F1, 266B2, 454C11, 33F8, 317G5, 520C9, and 260F-9-1C9, as determined by immunoprecipitation or sandwich immunoassay.

2. The monoclonal antibody of claim 1 wherein said reference antibody is selected from the group consisting of 260F9, 266B2, 113F1, 454C11 and 317G5.

3. A murine x murine hybridoma cell line that produces the monoclonal antibody of claim 2.

4. An immunotoxin comprising a conjugate of (a) the monoclonal antibody of claim 2; and (b) a cytotoxic moiety.

5. The monoclonal antibody of claim 2, wherein said antibody is labeled with a detectable label.

6. A method of diagnosing whether a human cell is a breast cancer cell comprising

(a) incubating a human cell with the antibody of claim 5; and

(b) determining the presence of labeled binary immune complexes on the human cell.

7. A method of diagnosing whether a human cell is a breast cancer cell comprising:

(a) incubating said human cell with the monoclonal antibody of claim 2;

(b) incubating the human cell with a labeled antibody against said monoclonal antibody;

(c) determining the presence of labeled ternary immune complexes on the human cell.

8. The monoclonal antibody of claim 1 wherein said reference antibody is 260F9 or 266B2.

9. The monoclonal antibody of claim 1 wherein said reference antibody is 454C11.

10. The monoclonal antibody of claim 9 wherein said antibody is labeled with a detectable label.

11. A method of diagnosing whether a human cell is a breast cancer cell comprising

(a) incubating a human cell with the antibody of claim 10; and

(b) determining the presence of labeled binary immune complexes on the human cell.

12. A method of diagnosing whether a human cell is a breast cancer cell comprising:

(a) incubating said human cell with the monoclonal antibody of claim 9;

(b) incubating the human cell with a labeled antibody against said monoclonal antibody;

(c) determining the presence of labeled ternary immune complexes on the human cell.

13. The monoclonal antibody of claim 1 wherein said reference antibody is 317G5.

14. The monoclonal antibody of claim 1 wherein said reference antibody is 113F1.

15. The monoclonal antibody of claim 1 wherein said reference antibody is 33F8.

16. The monoclonal antibody of claim 1 that binds the same epitope as the reference antibody.

17. The monoclonal antibody of claim 1 that is 260F9.

18. The cell line of claim 17 selected from the group consisting of 260F9, 113F1, 266B2, 454C11, 33F8, 317G5, 520C9, and 260F91C9.

19. The monoclonal antibody of claim 1 that is 260F9-1C9.

20. The monoclonal antibody of claim 1 wherein the exhibited TCID 50% is less than about 1 nM.

21. A murine x murine hybridoma cell line that produces the monoclonal antibody of claim 1.

22. An immunotoxin comprising a conjugate of

(a) the monoclonal antibody of claim 1 and

(b) a cytotoxic moiety.

23. The immunotoxin of claim 22 wherein the cytotoxic moiety is ricin A chain, PAPII, abrin A chain or a nonbinding, active fragment of diphtheria toxin.

24. A method of killing human breast cancer cells comprising contacting said cells with a cytocidally effective amount of the immunotoxin of claim 23.

25. The immunotoxin of claim 22 wherein the cytotoxic moiety is ricin A chain.

26. A method of killing human breast cancer cells comprising contacting said cells with a cytocidally effective amount of the immunotoxin of claim 22.

27. The monoclonal antibody of claim 1 wherein said antibody is labeled with a detectable label.

28. A method of diagnosing whether a human cell is a breast cancer cell comprising

(a) incubating a human cell with the antibody of claim 27 and

(b) determining the presence of labeled binary immune complexes on the human cell.

29. A method of diagnosing whether a human cell is a breast cancer cell comprising:

(a) incubating said human cell with the monoclonal antibody of claim 1;

(b) incubating the human cell with a labeled antibody against said monoclonal antibody;

(c) determining the presence of labeled ternary immune complexes on the human cell.

Cancer treatment method

Abstract
A treatment of cancer by the application of external electromagnetic energy capable of the generation of heat in intracellular particles to induce selective thermal death of cancer cells in living tissue. This process allows for the selective treatment of cancer cells in living tissue without damaging the normal cells.

Patent number: 4106488
Filing date: Jan 22, 1976
Issue date: Aug 15, 1978
Inventor: Robert Thomas Gordon

download


What is claimed is:

1. The process for the treatment of cancer cells without substantially damaging living normal cells comprising:

providing one of a tumor specific material selected from an antibody and a radioisotope,

encapsulating said tumor specific material with material capable of being removed upon entering the cancer cell after a preselected time, to produce a total particle size of less than 1 micron,

intravenously injecting the encapsulated particles of tumor specific material,

removing said encapsulating material, and

releasing said tumor specific material within the cancer cells.

2. The process of claim 1 including removing the encapsulating material by subjecting the patient to an alternating electromagnetic field to inductively heat the particle and melt the encapsulating material.

3. The process of claim 1 including removing the encapsulating material by subjecting the patient to an alternating electromagnetic field to vibrate the encapsulating material and destroy the integrity of the encapsulates.

4. The process of claim 1 including disolving the encapsulating material intracellularly.

5. A process for the treatment of cancer cells by application of external electromagnetic energy capable of the generation of heat in intracellular particles to induce selective thermal death of cancer cells comprising:

intravenously injecting into the patient minute particles capable of being inductively heated and of a size capable of being absorbed into the cancer cells,

absorbing said minute particles intracellularly into the cancer cells,

subjecting the patient to an alternating electromagnetic field to inductively heat the minute particles and thereby the cancer cells,

continuing the inductive heating of said particles to attain an increase in intracellular temperature to selectively kill the cancer cells.

6. A process for the treatment of cancer cells in living tissue by application of external electromagnetic energy capable of the generation of heat intracellularly to induce selective thermal death of the cancer cells comprising:

intravenously injecting into the patient minute particles capable of being inductively heated and of size less than 1 micron,

selectively absorbing said minute particles, intracellularly into the cancer cells,

subjecting the patient to an alternating electromagnetic field to inductively heat the minute particles and thereby the cancer cells,

continuing the inductive heating of said particles to attain an increase in intracellular temperature of at least 8.0.degree. Centigrade and not greater than 9.5.degree. Centigrade to kill the cancer cells.

7. The process of claim 1 wherein the particles are ferromagnetic, paramagnetic, or diamagnetic.

8. The process of claim 1 wherein the particles are selected from ferric hydroxide and iron oxide.

9. The process of claim 1 including providing a cancer cell seeking agent in a concentration sufficient to combine with and selectively direct the particles to the cancer cell.

10. The process of claim 9 wherein the cancer cell seeking agent is radioisotope selected from gallium-67, indium-113m, technetium-99m, fluorine and selenium-75.

11. The process of claim 9 wherein said cancer cell seeking agent is a tumor specific cancer antibody.

12. The process of claim 9 wherein the cancer cell seeking agent is gallium-67.

13. The process of claim 1 including forming the particles as a coating around a chemotherapeutic agent specific for treating cancer.

14. The process of claim 13 wherein the chemotherapeutic agent is selected from 5-flurouracil, nitrogen mustard, actinomycin D, methotrexate, cytoxan and vincristine.

15. The process of claim 13 including removing the coating of the cancer cell seeking agent after selectively directing the particles to the cancer cell.

16. The process of claim 13 including releasing the chemotherapeutic agent by removing the coating of said particles by subjecting the particles to an alternating electromagnetic field.

17. The process of claim 13 wherein the chemotherapeutic agent is nitrogen mustard.

Anti-cancer device

Abstract
An anti-cancer device having an anti-cancer drug and a blood coagulation factor fixed to a structure. This anti-cancer device is used in transcatheter arterial embolization and needle therapy with advantage and slowly releases the anti-cancer drug over an extended period by staying in the cancer tissue and its nearby area.

Patent number: 4536387
Filing date: Feb 14, 1983
Issue date: Aug 20, 1985
Inventors: Izumi Sakamoto, Kunihiko Takagi
Assignee: Unitika Ltd.

download


What is claimed is:

1. A method of transcatheter arterial embolization and needle therapy comprising injecting a patient with composition useful for transcatheter arterial embolization and needle therapy comprising:

(a) a polymer, wherein said polymer has fixed thereto;

(b) an anti-cancer drug; and

(c) a blood coagulation factor, wherein said anti-cancer drug and said blood coagulation factor are fixed to said polymer so as to be capable of sustained release from the polymer at the site of injection.

2. The method as claimed in claim 1, wherein said polymer is a synthetic polymer.

3. The method as claimed in claim 2, wherein said synthetic polymer is silicone.

4. The method as claimed in claim 1, wherein said polymer is selected from the group consisting of cellulosic material, cellulosic material derivative and regenerated cellulose.

5. The method as claimed in claim 4, wherein said cellulosic material derivative is ethyl cellulose.

6. The method as claimed in claim 1, wherein said polymer is bioabsorbable material.

7. The method as claimed in claim 6, wherein said bioabsorbable material is polysaccharide.

8. The method as claimed in claim 7, wherein said polysaccharide is amylose.

9. The method as claimed in claim 7, wherein said polysaccharide is oxidized cellulose.

10. The method as claimed in claim 7, wherein said polysaccharide is chitin.

11. The method as claimed in claim 6, wherein said bioabsorbable material is collagen.

12. The method as claimed in claim 6, wherein said bioabsorbable material is gelatin.

13. The method as claimed in claim 6, wherein said bioabsorbable material is polyamino acid.

14. The method as claimed in claim 13, wherein said polyamino acid is polyglycolic acid.

15. The method as claimed in claim 13, wherein said polyamino acid is polylactic acid.

16. The method as claimed in claim 1, wherein the polymer is in the form of a fibrous assembly.

17. The method as claimed in claim 16, wherein said fibrous assembly is an assembly of oxidized cellulose fiber.

18. The method as claimed in claim 16, wherein said fibrous assembly is an assembly of gelatin fiber.

19. The method as claimed in claim 16, wherein said fibrous assembly is an assembly of chitin fiber.

20. The method as claimed in claim 1, wherein said polymer is in the form of a sponge.

21. The method as claimed in claim 20, wherein said sponge is oxidized cellulose sponge.

22. The method as claimed in claim 20, wherein said sponge is gelatin sponge.

23. The method as claimed in claim 20, wherein said sponge is chitin sponge.

24. The method as claimed in claim 1, wherein said polymer is in the form of a powder.

25. The method as claimed in claim 24, wherein said powder is oxidized cellulose powder.

26. The method as claimed in claim 24, wherein said powder is gelatin powder.

27. The method as claimed in claim 24, wherein said powder is chitin powder.

28. The method as claimed in claim 1, wherein said polymer is in the form of a monofilament.

29. The method as claimed in claim 1, wherein said polymer is in the form of a film.

30. The method as claimed in claim 1, wherein said polymer is in the form of a microcapsule.

31. The method as claimed in claim 1, wherein said anti-cancer drug is an alkylating agent.

32. The method as claimed in claim 1, wherein said anti-cancer drug is a combination of cyclophosphamide, 5-fluorouracil and mitomycin.

33. The method as claimed in claim 1, wherein said anti-cancer drug is a combination of cyclophosphamide, 5-fluorouracil and bleomycin.

34. The method as claimed in claim 1, wherein said anti-cancer drug is bleomycin.

35. The method as claimed in claim 1, wherein said anti-cancer drug is mitomycin C.

36. The method as claimed in claim 1, wherein said anti-cancer drug is adriamycin.

37. The method as claimed in claim 1, wherein said anti-cancer drug is 5-fluorouracil.

38. The method as claimed in claim 1, wherein said blood coagulation factor is Factor XIII.

39. The method as claimed in claim 1, wherein said blood coagulation factor is thrombin.

40. The method as claimed in claim 1, wherein said blood coagulation factor is a combination of Factor XIII and thrombin.

41. The method as claimed in claim 1, wherein said anti-cancer drug and blood coagulation factor are fixed to the polymer by covalently bonding to the polymer.

42. The method as claimed in claims 1, 6, 16, 20, 24, 28, 29 or 30, wherein said anti-cancer drug and blood coagulation factor are fixed to the said polymer by ionically bonding to said polymer.

43. The method as claimed in claims 1, 6, 16, 20, 24, 28, 29 or 30, wherein said anti-cancer drug and blood coagulation factor are fixed to said polymer by adsorption.

44. The method as claimed in claims 1, 6, 16, 20, 24, 28, 29 or 30, wherein said anti-cancer drug and blood coagulation factor are fixed to said polymer for entrapping.

45. The method as claimed in claim 1, wherein calcium ion is additionally fixed to said polymer.

46. The method as claimed in claim 1, wherein a pharmaceutical is additionally fixed to said polymer.

47. The method as claimed in claim 46, wherein said pharmaceutical is selected from the group consisting of protease inhibitors, plasma proteins, fibronectin, antiboitics, antivirals, sulfaniamides and anti-infectives.

48. The method as claimed in claim 1, wherein said polymer is a bioabsorbable polymer and wherein said polymer is in the form of a fibrous assembly.

49. The method as claimed in claim 1, wherein said polymer is a bioabsorbable material and wherein said polymer is in the form of a sponge.

50. The method as claimed in claim 1, wherein said polymer is a bioabsorbable material and wherein said polymer is in the form of a powder.

51. The method as claimed in claim 1, wherein said polymer is a bioabsorbable material and wherein said polymer is in the form of a monofilament.

52. The method as claimed in claim 1, wherein said polymer is a bioabsorbable material and wherein said polymer is in the form of a film.

53. The method as claimed in claim 1, wherein said polymer is a bioabsorbable material and wherein said polymer is in the form of a microcapsule.

54. The method as claimed in claim 1, wherein said polymer is gelatin powder, said anti-cancer drug, and said blood coagulation factor are fixed to said polymer by ionic bonding, entrapping and adsorption, said anti-cancer drug is mitomycin C and wherein said blood coagulation factor is a combination of Factor XIII and thrombin.

Cancer treatment by intracellular hyperthermia

Abstract

A treatment of cancer by the application of chemical reactions intracellularly capable of the intracellular generation of heat so as to induce selective thermal death of cancer cells in living tissue. Metabolizable minute particles of a size less than one micron are intravenously injected into the patient and absorbed by the cancer cells. The oxygen level of the patient's blood is then increased. The rate of intracellular chemical reaction in the cancer cells due at least in part to the intracellular presence of these minute particles is thereby increased and intracellular heat generated. The oxygen level is increased until the intracellular temperature has increased at least 8.0 degrees Centigrade but not more than 9.5 degrees Centigrade thereby selectively killing the cancer cells without damaging the normal cells.

Patent number: 4569836
Filing date: May 24, 1983
Issue date: Feb 11, 1986
Inventor: Robert T. Gordon

download


What is claimed is:

1. A process for the treatment of cancer cells in living tissue of a patient comprising the following steps:

introducing into the living tissue of the patient minute particles of a size not greater than one micron and capable of being metabolized by the cancer cells and capable of an increased rate of metabolism or oxidation by the increased availability of oxygen,

absorbing said minute particles intracellularly into the cancer cells, thereafter, increasing the oxygen level of the blood of the patient, and thereby increasing the rate of intracellular chemical reaction in the cancer cells due at least in part to the intracellular presence of said minute particles, and generating intracellular heat therein, and

continuing said increasing the oxygen level step until the intracellular temperature has increased at least 8.0 degrees Centigrade but not more than 9.5 degrees Centigrade to selectively induce thermal death of the cancer cells.

2. The process of claim 1 including,

said increasing the oxygen level step including subjecting the patient to hyperbaric air.

3. The process of claim 1 including,

said increasing the oxygen level step including subjecting the patient to hyperbaric oxygen.

4. The process of claim 1 including,

said increasing the oxygen level step including increasing the levels of 2,3-Diphosphoglycerate in the body to enhance the availability of the oxygen to the cells and thereby increasing the metabolism in the cells.

5. The process of claim 1 including,

said increasing the oxygen level step including introducing phosphates into the patient to increase the availability of oxygen to the cells and thereby increasing intracellular metabolism.

6. The process of claim 5 including,

said increasing the oxygen level step including subjecting the patient to hyperbaric oxygen.

7. The process of claim 1 including,

said minute particles comprising a compound which can be further oxidized.

8. The process of claim 7 including,

said compound being ferric oxyhydroxide, ferric hydroxide, iron carbonate, or iron citrate.

9. The process of claim 1 including,

said introducing step including intravenously injecting into the patient said minute particles.

10. The process of claim 1 including,

said introducing step including intravenously injecting into the patient said minute particles suspended in a liquid vehicle.

11. The process of claim 10 including,

said minute particles being integrated with a sugar molecule.

12. the process of claim 11 including,

said sugar molecule being dextrose, dextran, glucose or sucrose.

13. The process of claim 1 including,

said minute particles being bound to radioisotopes.

14. The process of claim 1 including,

said minute particles being bound to cancer antibodies.

15. The process of claim 1 including,

said minute particles comprising minute encapsulated chemotherapeutic agent particles.

16. The process of claim 1 including,

said minute particles comprising minute integrated chemotherapeutic agent particles.

17. The process of claim 1 including,

said minute particles comprising radioisotopes, and

said absorbing step including said radioisotopes being absorbed selectively in said cancer cells.

18. The process of claim 17 including,

said radioisotopes being gallium-67, indium-113m, technetium-99m, fluorine, or selenium-75.

19. The process of claim 1 including,

said introducing step including introducing a cancer cell seeking agent in a concentration sufficient to combine with and selectively direct said minute particles to the cancer cells.

20. The process of claim 19 including,

said cancer cell seeking agent being a radioisotope.

21. The process of claim 20 including,

said radioisotope being gallium-67, indium-113m, technetium-99m, fluorine or selenium-75.

22. The process of claim 19 including,

said cancer cell seeking agent being a tumor specific cancer antibody.

23. The process of claim 1 including,

said minute particles including a chemotherapeutic agent specific for treating cancer and a coating around said chemotherapeutic agent.

24. The process of claim 23 including,

said chemotherapeutic agent being 5-flurouracil, nitrogen mustard, actinomycin-D, methotrexate, cytoxan, or vincristine.

25. The process of claim 23 including,

removing said coating from said chemotherapeutic agent, after said absorbing step.

26. The process of claim 15 including,

said removing step including subjecting said minute particles to an increased oxygen supply.

27. The process of claim 23 including,

said increasing the oxygen level step including removing said coating from said chemotherapeutic agent, after said absorbing step.

28. A process for the treatment of cancer cells in living tissue of a patient comprising the following steps:

introducing into the living tissue of the patient minute particles of a total size not greater than one micron, capable of metabolizing and of reacting with another chemical, and having a chemotherapeutic agent and an agent coating solubilizable after a period of time by the cytoplasm of the cancer cell,

depositing said minute particles intracellularly into the cancer cells, after said introducing step, and

at least said period of time after said depositing step, solubilizing said coating and absorbing said chemotherapeutic agent in the cancer cells resulting in their death.

29. The process of claim 28 including,

said chemotherapeutic agent being nitrogen mustard, antinomycin D, methotrexate, 5-flurouracil, cytoxan, or vincristine, and

said coating comprising an iron dextran complex material.

30. The process of claim 29 including,

after said depositing step before said period of time has passed, subjecting said minute particles to an increased oxygen supply to remove said coating.

Methods and compositions for treatment of cancer using oligonucleotides

Abstract

The present invention provides methods useful in autologous bone marrow transplantation and cancer therapy. According to one aspect of the invention, bone marrow cells from a patient having cancer are treated with selected antisense oligonucleotides in order to deplete the bone marrow of malignant cells prior to infusion back into the bone marrow donor. In a separate embodiment, selected antisense oligonucleotides are administered systemically for anticancer therapy.

Patent number: 5087617
Filing date: Feb 15, 1989
Issue date: Feb 11, 1992
Inventor: Larry J. Smith
Assignee: Board of Regents, The University of Texas System

International Classification
A61K 3170

download

What is claimed is:

1. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:

a obtaining bone marrow cells from the individual; and

b exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of those genes encoding cell surface receptors, modulators of intracellular messengers, or transcriptional regulators.

2. A method for treating an individual having cancer comprising the steps of:

a obtaining bone marrow cells from the individual; and

b exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of those genes encoding cell surface receptors, modulators of intracellular messengers, or transcriptional regulators; and

c transplanting the exposed cells into the individual.

3. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:

a) obtaining bone marrow cells from the individual; and

b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of genes that encode a molecule that regulates cell proliferation or viability.

4. A method for treating an individual having cancer comprising the steps of:

a) obtaining bone marrow cells from the individual; and

b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of genes that encode a molecule that regulates cell proliferation or viability.

c) transplanting the exposed cells into the individual.

5. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:

a) obtaining bone marrow cells from the individual; and

b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is a gene that encodes a molecule that regulates cellular differentiation.

6. A method for treating an individual having cancer comprising the steps of:

a) obtaining bone marrow cells from the individual; and

b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is a gene that encodes a molecule that regulates cellular differentiation; and

c) transplanting the exposed cells into the individual.

7. The method of claim 1, 2, 3, 4, 5 or 6 wherein said oligonucleotide has sufficient complementarity with the target gene to form a duplex having a melting temperature of at least about 40 degrees Centigrade under physiologic conditions.

8. The method of claim 2 or 4 or 6 comprising the additional step of administering to the individual a sufficient amount of a preparation containing oligonucleotides complementary to RNA transcribed from a target gene present in the cells of the cancer to kill the cancerous cells of the individual.

9. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is further defined as a cancer of cells of the hemopoietic system.

10. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is leukemia.

11. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is myeloid leukemia.

12. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is lymphoma.

13. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is breast cancer.

14. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is multiple myeloma.

15. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is gastrointestinal cancer.

16. The method of claim 1, 2, 3, 4, 5 or 6 where the target is a traitor gene.

17. The method of claim 1, 2, 3, 4, 5 or 6 wherein the oligonucleotide has a charged backbone.

18. The method of claim 1, 2, 3, 4, 5 or 6 wherein the oligonucleotide has an uncharged backbone.

19. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide is a methylphosphonate oligonucleotide.

20. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide is a phosphorothioate oligonucleotide.

21. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide comprises at least 8 bases and is complementary to a sequence of RNA located 5' to the initiation condon of said target.

22. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide comprises the sequence 3'-GGTCTGACGGAAGGCCCAGTGACGGTAC-5'.

23. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide comprises at least 8 bases and is located within 40 bases of the initiation codon.

24. The method of claim 1, 2, 3, 4, 5 or 6 where the proliferation-inhibiting amount is 10-200 micromolar.

25. The method of claim 1, 2, 3, 4, 5 or 6 where the exposing step comprises:

a. fractionating the bone marrow cells to obtain a mononuclear cell fraction, and

b. culturing the mononuclear fraction together with a proliferation inhibiting amount of the oligonucleotide for at least about 1-10 days.

26. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:

a) obtaining bone marrow cells from the individual;

b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is p53.

Device for treating cancer and non-malignant tumors and methods

Abstract
A method of this invention for treating body tissues containing cancerous cells or non-malignant tumors with RF ablation, alone or in combination with systemic or localized chemotherapy comprising introducing a stylet comprising an electrode surface and a sleeve longitudinally moveable thereon into the vicinity of the body tissues, retracting the sleeve from a portion of the electrode surface, and supplying RF power to the electrode surface sufficient to heat the tissue to a temperature of above about 45.degree. C. for a time to cause reduction of tissue mass in the vicinity of the electrode. The RF power supplied to the electrode surface is sufficient to effect a desiccated fluid diffusion barrier capsule surrounding the body tissue being treated. The stylet can include a hollow tube having fluid distribution ports therein, and the method can include the step of passing fluid through one or more distribution ports into the body tissue being treated. The fluid can be saline...

Patent number: 5472441
Filing date: Mar 11, 1994
Issue date: Dec 5, 1995
Inventors: Stuart D. Edwards, Ronald G. Lax
Assignee: Zomed International
Primary Examiner: Michael Peffley

download

What is claimed is:

1. A method for ablating tissue at a tissue treatment site where dehydration of tissue at the tissue treatment site is reduced, the method comprising the steps of:

a) introducing an Rf ablation device into a tissue treatment site, the device including a hollow tubular Rf electrode having a closed, sharpened distal tip, an electrode conductive surface for conducting Rf ablation, and a plurality of fluid distribution ports distributed along the length of the electrode for delivering chemotherapeutic agents to the tissue treatment site; and

b) supplying Rf power to the electrode to produce a diffusion barrier capsule at the tissue treatment site wherein the position of the electrode relative to the tissue treatment site is varied sufficiently frequently to reduce dehydration of tissue at the tissue treatment site.

2. The method according to claim 1, further including the step of passing fluid through the fluid distribution ports during ablation to reduce dehydration of tissue at the tissue treatment site.

3. The method according to claim 1 further including the step of varying the position of the electrode relative to the tissue treatment site by at least 1 mm during ablation.

4. The method according to claim 2 further including the step of varying the position of the electrode relative to the tissue treatment site between about 1 and 3 mm.

5. The method according to claim 1 further including the step of varying the position of the electrode relative to the tissue treatment site along a longitudinal axis of the electrode.

6. The method according to claim 1 wherein the device further includes an electrode position modifier and further including the step of varying the position of the electrode relative to the tissue treatment site sufficiently frequently during ablation to reduce dehydration of tissue at the tissue treatment site.

7. The method according to claim 5 further including the step of passing fluid through the fluid distribution ports during ablation to reduce dehydration of tissue at the tissue treatment site.

8. The method according to claim 7 further including the step of varying the position of the electrode relative to the tissue treatment site is by at least 1 mm during ablation.

9. The method according to claim 8 further including the step of varying the position of the electrode relative to the tissue treatment site by between about 1 and 3 mm.

download pdf :